WO2016068600A1 - Composition pour un traitement visant les cellules souches cancéreuses - Google Patents

Composition pour un traitement visant les cellules souches cancéreuses Download PDF

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WO2016068600A1
WO2016068600A1 PCT/KR2015/011436 KR2015011436W WO2016068600A1 WO 2016068600 A1 WO2016068600 A1 WO 2016068600A1 KR 2015011436 W KR2015011436 W KR 2015011436W WO 2016068600 A1 WO2016068600 A1 WO 2016068600A1
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cancer
stem cells
composition
inhibitor
cancer stem
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PCT/KR2015/011436
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English (en)
Korean (ko)
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정재호
박은성
박기청
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연세대학교 산학협력단
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Priority to JP2017523451A priority Critical patent/JP6446134B2/ja
Priority to DK15856097.9T priority patent/DK3213752T3/da
Priority to CN201580071381.5A priority patent/CN107106580B/zh
Priority to US15/522,558 priority patent/US10420814B2/en
Priority to EP15856097.9A priority patent/EP3213752B1/fr
Priority to ES15856097T priority patent/ES2743493T3/es
Publication of WO2016068600A1 publication Critical patent/WO2016068600A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7004—Monosaccharides having only carbon, hydrogen and oxygen atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/351—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom not condensed with another ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4422—1,4-Dihydropyridines, e.g. nifedipine, nicardipine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/455—Nicotinic acids, e.g. niacin; Derivatives thereof, e.g. esters, amides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00

Definitions

  • the present invention relates to a composition for treating cancer stem cells, comprising a glucose uptake inhibitor (hico tor) and a cal cium pump inhibitor (Ib i tor).
  • cancer stem cells are cancer cells with unlimited regenerative capacity similar to those of general stem cells, and proliferate slowly differently from general cancer cells, and are known as cancer cells having self-renewal or differentiation ability, which is a characteristic of enjoyable cells.
  • mechani sm there is no active research on cancer stem cells, and there is almost no research on drugs for treating cancer-producing cells that specifically target cancer stem cells.
  • composition for treating cancer enjoyment cells effective for cancer stem cells is expected to be an effective treatment method that can not only increase the therapeutic effect of cancer but also suppress the recurrence and / or metastasis of cancer.
  • the present invention has been made to solve the above problems in the prior art, to provide a composition for treating cancer stem cells comprising a glucose uptake inhibitor and a calcium pump inhibitor (cal cium pump inhibi tor) For that purpose.
  • cancer stem cel l means a cancer cell having a comprehensive meaning of self-renewal and differentiation ability, which is a characteristic unique to stem cells.
  • the cancer stem cell is a normal tumor growth condition (the "normal tumor growth condition” refers to a state in which there is not enough cellular nutrients (glucose) necessary for cell growth and abundant growth conditions of the tumor microenvironment.
  • the present invention provides a composition for inhibiting cancer stem cell growth or a pharmaceutical composition for treating cancer stem cells, comprising a glucose uptake inhibitor and a calcium pump inhibitor as an active ingredient.
  • the glucose absorption inhibitor is preferably a glucose derivative (glucose der ivat ive), more preferably 2-deoxyglucose (2 ⁇ deoxyglucose, 2DG) or, the absorption of sugar which is the energy source of the cell Restricts cell growth by inducing nutrient deprivation conditions and / or metabolic energy depletion-associated vesicle stress states, and cancer.
  • a glucose derivative glucose derivative
  • Any compound that induces the expression of pl asma membrane Ca 2+ ATPase (PMCA) in stem cells is not limited thereto.
  • the derivatives refer to analogous compounds obtained by changing a part of glucose, and compounds which act competitively with normal glucose to inhibit the absorption of
  • the inhibitor of PMCA means a substance capable of inhibiting the release of the scab to the outside of the cell by inhibiting the activity of PMCA.
  • the composition may further comprise a biguamde-based compound.
  • the biguanide-based compound is preferably a biguanide-based diabetic therapeutic agent, more preferably metformin (met formin), phenformin, buformine, but hinders energy production in the cell. If it is a biguanide-based compound that induces a nutritional deficiency-like state is not limited thereto.
  • the cancer is preferably breast cancer, uterine cancer, stomach cancer, brain cancer, rectal cancer, colon cancer, lung cancer, skin cancer, ovarian cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, thyroid cancer, liver cancer, etc. It may be, and more preferably may be breast cancer, but the cancer progression such as tumor differentiation and / or proliferation is not limited to the type of cancer depending on the cancer stem cells described in the present invention.
  • the cancer stem cells pharmaceutical composition for the treatment additionally other anticancer agents and in combination may be administered, may be used to treat them to effective cancer stem cells as well as the general cancer over, washing-based pharmaceutical composition inhibiting the recurrence of or all of the cancer, It can also be used as a pharmaceutical composition.
  • the pharmaceutical composition may be characterized in that the capsule, tablets, granules, injections, ointments, powder or beverage form, the pharmaceutical composition may be characterized in that it is intended for humans.
  • the pharmaceutical composition is not limited thereto, but may be used in the form of powder, granules, capsules, tablets, oral dosage forms such as aqueous suspensions, external preparation suppositories, and sterile injectable solutions according to conventional methods.
  • the pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers may be a binder, suspending agent, disintegrating agent 1, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, flavoring agents, etc.
  • compositions of the present invention can be prepared in various ways in combination with the pharmaceutically acceptable carriers described above.
  • oral administration may be in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc. It may be prepared in a dosage ampule or multiple dosage forms. Others may be formulated into solutions, suspensions, tablets, capsules, sustained release preparations and the like.
  • suitable carriers, excipients and diluents suitable for the formulation include lactose, dextrose, sucrose, sorbetle, manny, xylly, erythri, malditol, starch, acacia rubber, alginate, gelatin, calcium Phosphate, stump silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil can be used.
  • it may further include fillers, anti-fog agents, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like.
  • the route of administration of the pharmaceutical composition according to the present invention is not limited thereto, but is oral, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, transdermal, physiological, intraperitoneal, intranasal, intestinal, topical Includes sublingual or rectal. Oral or parenteral release is preferred.
  • parenteral includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intramuscular, sternum, intradural, intralesional and intracranial injection or infusion techniques.
  • the composition may also be administered in the form of suppositories for rectal administration.
  • the composition activity of the specific compound employed, the age, body weight, general -3 ⁇ 4 Kim ⁇ "sex-specific information ⁇ - wherein administration Guo-glass administration - ⁇ eu drug release rate of a hard and 7 ⁇ yo-times the sum o —And—prevention—r may vary depending on a number of factors, including the severity of the particular disease, and the dosage of the pharmaceutical composition may vary from the patient's condition, weight, degree of disease, drug form, Depending on the route and duration of administration, it may be appropriately selected by those skilled in the art and may be administered at 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day.
  • Administration may be administered once a day or may be administered in divided several times i.
  • the dosage does not limit the scope of the invention in any aspect.
  • the pharmaceutical composition according to the present invention can be formulated as pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions.
  • a composition for treating cancer stem cells comprising a glucose uptake inhibitor and a calcium pump inhibitor, according to the present invention
  • Glucose absorption inhibitors and additional biguanide-based drugs are used to induce vesicle stress conditions related to nutritional deficiency and metabolic energy depletion in cancer stem cells, thereby inducing PMCA expression in cancer stem cells
  • a pump inhibitor in combination, cancer stem cells lower resistance to Ca 2+ -associated cell death and induce cancer cell death, which can be used as an effective cancer enjoyment cell therapy, thereby effectively treating various cancer stem cells. It is expected to be able to effectively suppress cancer recurrence and / or metastasis.
  • FIG. 1 is a diagram showing the results of confirming the survival rate in the glucose deficient state of the s-MM-MB-231 and s_MCF_7 cell line by crystal vitreous staining method according to an embodiment of the present invention.
  • FIG. 2 is a view showing the results of confirming the survival rate in the glucose deficient state s-MDA-MB-231 and s- MCF-7 cell line according to an embodiment of the present invention by ⁇ assay.
  • FIG. 3 is a diagram showing the TUNEL assay results according to an embodiment of the present invention.
  • 4 is a diagram showing the results of immunological analysis according to an embodiment of the present invention.
  • 5 is a view showing a cell cycle analysis results according to an embodiment of the present invention.
  • 6 is in accordance with an embodiment of the present invention. Figure showing the results of confirming the amount of Ca 2+ in the cells.
  • FIG. 7 is a view showing the results of confirming the expression level of CaMK-2 a according to an embodiment of the present invention.
  • FIG. 8 is a diagram showing the result of confirming the amount of Ca 2+ in the cell line in which the expression of _CaMK-2 a is suppressed according to an embodiment of the present invention.
  • FIG. 9 is a diagram showing the results of TUNEL assay in the cell line suppressed the expression of CaMK-2a according to an embodiment of the present invention.
  • FIG. 10 is a diagram showing the results of immunological analysis in a cell line in which the expression of CaMK-2 a is suppressed according to an embodiment of the present invention.
  • FIG. 12 is a diagram illustrating a result of confirming a correlation between PGC-1 a and 0 (—21) according to an embodiment of the present invention.
  • FIG. 13 is a diagram showing the results of confirming the amount of intracellular Ca 2+ in a cell line in which PGC-1 a expression is suppressed according to an embodiment of the present invention.
  • FIG. 14 is a diagram illustrating a result of confirming a binding relationship between PGC—la and PMCA1 by EMSA according to an embodiment of the present invention.
  • FIG. 15 is a diagram illustrating a result of confirming a binding relationship between PGC-1 a and PMCA2 by EMSA according to an embodiment of the present invention.
  • 16 is a view showing the results of confirming the amount of PMCA expression in a nutritional deficient state according to an embodiment of the present invention.
  • 17 is an embodiment of the present invention. Shows the results of confirming the amount of PMCA expression in the cell line suppressed the expression of CaMK-2a.
  • FIG. 19 is a diagram showing the results of confirming the protein expression by immunochemical staining in a breast cancer animal model according to one embodiment of the present invention.
  • 20 is a view showing the results of confirming the anticancer effect through the combined administration in the animal model of breast cancer according to an embodiment of the present invention.
  • 21 is a view showing the results of confirming the difference between cancer cells and cancer stem cells and gene expression according to an embodiment of the present invention.
  • FIG. 22 is a view showing the results confirmed by Western blotting the expression of calcium regulatory proteins of cancer cells and cancer stem cells according to an embodiment of the present invention.
  • Figure 23 is a schematic diagram showing a brief control of the ion control mechanism of cancer stem cells according to an embodiment of the present invention.
  • 24 is a view showing the results of confirming the change in CaMK-2a signaling mechanism with time according to an embodiment of the present invention.
  • 25 is a view showing the results of confirming the role of p NF—kB according to an embodiment of the present invention.
  • 26 is a view showing the TUNEL assay results according to an embodiment of the present invention.
  • 27 is a view showing the results of Western blotting according to an embodiment of the present invention.
  • 28 is a view showing the results of confirming the survival rate of the cells according to an embodiment of the present invention.
  • 29 is a view showing the results of confirming the growth of cancer in an animal model based on cancer stem cells according to an embodiment of the present invention.
  • FIG. 30 is a view showing the results of confirming the protein expression of the animal model based on cancer stem cells according to an embodiment of the present invention.
  • 31 is a view showing the results of confirming the protein expression of the animal model based on cancer stem cells according to an embodiment of the present invention.
  • cancer stem cells To prepare cancer stem cells, the parental cells of breast cancer cell lines MDA—MB-231 and MCF-7 cell lines, p_MDA-MB_231 and p-MCF_7, induce long-term nutritional deficiency. Cancer stem cells were evacuated from nutrient deprivation and surviving cells (s-MDA-MB-231 and s-MCF—7) were screened and tested through conventional cancer stem cell specific biological characteristics to produce cancer stem cells. -. In order to elucidate the mechanism of cancer stem cells and to develop a therapeutic agent that can inhibit cancer enjoyment cells, s-MDA-MB-231 and s-MCF-7 cells were transfected with shPGC-lapGFP-V-RS vector (Origene), respectively.
  • shPGC-lapGFP-V-RS vector Origene
  • sshPGC-la-MDA-MB-231 and s-shPGC-1 a -MCF-7 which are stem cells stably expressing shPGOla, were prepared and used for the experiment.
  • Each cell line contains 10% fetal bovine serum (FBS). Cultured in the included RPMI-1640 medium.
  • P-MDA-MB-231, p-MCF-7, s-MDA-MB- prepared in the same manner as in Example 1 to compare the survival rate of cancer cells and cancer stem cells in a glucose depr ivat ion. 231, and s-MCF-7 cell lines were added to 96 well plates at a concentration of 5 ⁇ 10 3/100 uL, respectively, and cultured until about 70% of the culture vessel area, followed by the addition of 10% FBS deficient in glucose. was further cultured for 3 days by exchange with RPMI-1640 medium. And cell survival was confirmed at 0, 12, 24, 36, 48, 60, and 72 hours, respectively.
  • cancer stem cells s-MDA-MB-231 and s—MCF-7 cell line, were the parent cell p. -Compared to MDA-MB-231 and p-MCF-7 . -It was confirmed that it showed a very high freshness rate.
  • the early stage of culture (after 12 hours of cultivation) does not show a significant difference in the survival rate between cancer stem cells and general cancer cells, but as the late stage of culture (after 48 hours of cultivation) It was confirmed that there is a significant difference in the survival rate with general cancer cells.
  • the survival rate between cancer cells and cancer stem cells is more than 40%.
  • p-MDA-MB—231, p—MCF-7, s-MDA-MB-231, and s-MCF—7 cell lines were recovered and subjected to immunological analysis.
  • the recovered cells were washed twice with cold PBS (phosphate buffered saline) buffer, and proteins were isolated by lysing the cells with RIPA buffer solution.
  • the isolated protein was analyzed by BCA- assay for further experiments. The amount of was measured.
  • cancer stem cells have high resistance to apoptosis (ant i-apoptos i s) compared to general cancer cells in nutrient deprivation state and thus show high survival even in nutrient deprivation state.
  • Example 4 Identification of Causes of Apoptosis of Cancer Stem Cells
  • CaMK-2 as iR A was transfected (t ransectect on) to produce cancer stem cell lines with inhibited CaMK-2 ⁇ expression. And the amount of CaMK-2 ex and the amount of Ca 2+ in the cells were measured in the same manner as in Example 4. 1 and 4.2. The results are shown in FIG.
  • Example 3 TUNEL assay, immunological analysis, and in the same manner as in Example 3 using cancer stem cell lines suppressed CaMK-2 ⁇ expression. Cell cycle analysis was performed. The results are shown in FIGS. 9 to 11, respectively.
  • cancer stem cells generally exhibit resistance to apoptosis due to malnutrition, but in the case of cancer stem cells which inhibit the expression of CaMK-2 ⁇ using s iRNA, common cancer cells Likewise, apoptosis was induced and DNA fragmentation was induced.
  • cancer stem cells for increasing the expression level of CaMK-2a in a nutrient deficiency by adjusting the amount of Ca 2+ inside cells to an appropriate level Ca 2+ associated cell death (apoptosis mediated Ca 2+) Resistant, and inhibiting the expression of CaMK-2 ⁇ in cancer stem cells was confirmed to lose the resistance.
  • PGC-1 a peroxisome prol iferator activated receptor gamma (; 03 (: 1; ⁇ 01 " 1)) and ⁇ -201 association
  • the sh PGC-la vector in the s ⁇ MDA-MB-231 cell line ( Oi'igene) was transfected (transfect ion) to produce a cancer stem cell line that inhibited the expression of PGOla, ' immunological analysis was carried out in the same manner as in Example 3.2. The results are shown in FIG.
  • EMSA assay was performed according to the protocol using the EMSA kit.
  • binding sequence "TTGACCTTTGGCCCA 1 ', which is the binding site sequence of the promoter of PMCA1, was used. The results are shown in FIG.
  • EMSA assay was performed according to the protocol using the EMSA kit.
  • -As the binding sequence the "CTGG ATACCCC” which is the binding site sequence of the promoter of PMCA2 is used.
  • CTGG ATACCCC the binding site sequence of the promoter of PMCA2 is used.
  • NF-kB, PGC-1 ⁇ and DNA (probe) binding increased in nutrient deficient cancer stem cells, and ant i—p65 or ant i-p50, an antibody to NF-kB.
  • supershi ft occurred by additionally binding the antibody.
  • PGC-1 a binds to the promoter region of PMCA2 in combination with NF-kB and regulates the expression of PMCA2 in a malnourished state.
  • each cell line was cultured in the malnourished state in the same manner as in Example 2, and subjected to QRT ⁇ PCR according to the culture time.
  • Cells recovered at each time were extracted with RNA according to the protocol using RNeasy Mini Ki t, and QRT-PCR was performed using one step RT-PCR ki t using extracted RNA lug.
  • the primer sequences used are listed in Table 1. The results are shown in FIG. In addition, the results of the same experiment using the cell line suppressing the expression of ⁇ & «—2 (1) are shown in FIG. 17.
  • calcium pump inhibitor acts as an inhibitor of PMCA acting as Ca 2+ -ATPase in plasma membrane PMCA or thapsigargin, an inhibitor of sarco / endoplasmic reticulum Ca 2 + -ATPase, which acts as a Ca 2+ _ATPase in the endoplasmic reticulum.
  • SERCA activity After inhibiting SERCA activity, the change in extracellular Ca 2+ amount in cancer stem cells was measured. The results are shown in FIG. 18.
  • cancer stem cells in the nutrient deprivation state and metabolic energy depletion-related vesicle stress state increase the expression of PGC-1 ⁇ to promote the expression of CaMK-2a, and also act as a coactivator of PMCA1 and PMCA2, PMCA protein apoptosis is associated with an increased expression of the secreted from the cytoplasm to the endoplasmic reticulum in a nutrient deficient cell
  • Ca 2+ is Ca 2+ through the action of discharging to the outside to be stored in mitochondrial Ca 2+ induced apoptosis (Ca 2+ mediated apoptosis).
  • a breast cancer animal model was produced.
  • breast cancer cells p— MDA-MB-231 and p ⁇ MCF-7 cell lines and breast cancer stem cells s— MDA-MB— 231 and s-MCF-7 cell lines were cultured in vitro, respectively.
  • Breast cancer animals are injected into the upper left flank of BALB / c nude mice at week 5-6 to give 1.0X10 7 eel 1 s / mouse, provide water and feed for 7 days, and adjust light at 22 ° C for 12 hours.
  • the model was produced.
  • Inject inperi toneal inj ect ion
  • cancer stem cells hardly grew when 2—de-glucose, caloxin, and metformin were administered in combination. These results are in combination with glucose absorption inhibitors.
  • Use of biguanide drugs to induce nutritional deficiency and metabolic energy depletion-related vesicle stress in cancer stem cells, thereby inducing PMCA expression in cancer stem cells and co-administering pumps By lowering the resistance to Ca 2+ -cell killing of cancer stem cells by inducing cancer cell death, it was confirmed that it can be used as an effective cancer stem cell specific treatment method. It was confirmed that it is possible to effectively suppress the recurrence and / or metastasis caused by the cancer stem cells which is the limit.
  • Example 7 Confirmation of survival mechanism of cancer stem cells
  • Example 2 In order to confirm the survival mechanism of cancer stem cells, the same method as in Example 2 P-MDA-MB-231, p-MCF-7, s-MDA-MB-231, and s-MCF-7 cell lines incubated for 40 hours in a glucose deficient state were recovered, respectively, in the same manner as in Example 5.3. RNA was extracted and subjected to microarray (mi croan-ay). The results are shown in FIG.
  • Example 3.2 Western blotting was carried out in the same manner as in Example 3.2 to confirm the difference in expression of genes that regulate calcium. The results are shown in FIG. As shown in FIG. 22, it was confirmed that the expression of SERCA2 was increased in the cancer stem cells, whereas the expression of IP3R was decreased. In general, when glucose is insufficient, the cells secrete calcium ions into the cytoplasm through the pathway called IP3R, and when the secretion of the chestnut increases rapidly, finally cell death is induced. It is known that this reabsorption can suppress cell death.
  • p NF-kB acted as a transcr ipt ion factor that increases the expression of Be 1-2 and SERCA2, which are inhibitors of IP3R.
  • NF-kB is phosphorylated and activated, and the activated NF-kB increases the expression of Bcl-2 and CERCA2, which increases reabsorption of the scabbard and inhibits the expression of IP3R. It was confirmed that the secretion of ions was suppressed.
  • KN62 L— [ ⁇ , 0-bis (5-i soqu i no 1 1 nesu 1 phony 1), well known as an inhibitor of CaMK-2 ⁇ -N-methyl-L-tyrosyl] -4-phenylpiperaz ine) was treated with cancer cells at a concentration of 10 uM, followed by TUNEL assay in the same manner as in Example 3. 1 to confirm cell death and DNA fragmentation.
  • Western blotting was performed in the same manner as in Example 3.2 to confirm the expression of genes, and the survival of cells was confirmed in the same manner as in Example 2. The results are shown in FIGS. 26 to 28.
  • Example 6 In order to confirm the characteristics of the animal model produced in the same manner as in Example 6, the resulting breast cancer tissue was extracted to measure the size of the cancer. The result is shown in FIG. Indicated. In addition, of breast cancer tissue. The degree of protein expression was confirmed by Western blotting and immunochemical staining. Western blotting was performed in the same manner as in Example 3.2, and immunochemical staining was performed in the same manner as in Example 6.2. The results are shown in FIGS. 30 and 31.
  • the animal model based on cancer stem cells was confirmed that the growth rate of cancer is faster than the animal model based on normal cancer cells.
  • the expression of IP3R was decreased in comparison with the animal model based on cancer cells, and the SERCA2 It was confirmed that expression was increased.
  • cancer stem cells activate CaMK-2a signaling mechanisms in poor conditions in the body.
  • expression of IP3R is suppressed and expression of SERCA2 is increased, resulting in cell death due to the release of calcium ions. It was confirmed that it can increase the viability of cancer in the body and promote recurrence and / or metastasis.
  • the composition for treating cancer stem cells effective for cancer stem cells is an effective treatment method that can not only maximize the therapeutic effect of cancer by overcoming the limitations of existing cancer treatment, but also suppress the recurrence and / or metastasis of cancer. It could be confirmed that it can be used.
  • composition of the present invention can be used as a therapeutic agent for cancer stem cells by effectively inducing the death of cancer stem cells, thereby effectively treating various cancer stem cells to effectively inhibit the recurrence and / or metastasis of cancer It can be used as a composition.

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  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne une composition pour un traitement efficace visant les cellules souches cancéreuses et, plus particulièrement, une composition contenant un inhibiteur de la capture du glucose, un composé à base de biguanide, et un inhibiteur de la pompe à calcium. La composition de la présente invention peut être utilisée en tant qu'agent thérapeutique visant les cellules souches cancéreuses du fait qu'elle induit de façon efficace la mort des cellules souches cancéreuses permettant ainsi un traitement efficace visant diverses cellules souches cancéreuses, par conséquent, la composition de la présente invention est censée inhiber de manière efficace la récurrence et/ou la métastase du cancer.
PCT/KR2015/011436 2014-10-29 2015-10-28 Composition pour un traitement visant les cellules souches cancéreuses WO2016068600A1 (fr)

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JP2017523451A JP6446134B2 (ja) 2014-10-29 2015-10-28 癌幹細胞治療用組成物
DK15856097.9T DK3213752T3 (da) 2014-10-29 2015-10-28 Sammensætning til behandling af cancerstamceller
CN201580071381.5A CN107106580B (zh) 2014-10-29 2015-10-28 治疗癌症干细胞的组合物
US15/522,558 US10420814B2 (en) 2014-10-29 2015-10-28 Composition for treating cancer stem cells
EP15856097.9A EP3213752B1 (fr) 2014-10-29 2015-10-28 Composition pour un traitement visant les cellules souches cancéreuses
ES15856097T ES2743493T3 (es) 2014-10-29 2015-10-28 Composición para el tratamiento de células madre cancerosas

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KR20140147972 2014-10-29
KR10-2014-0147972 2014-10-29

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EP (1) EP3213752B1 (fr)
JP (1) JP6446134B2 (fr)
KR (2) KR101774652B1 (fr)
CN (1) CN107106580B (fr)
DK (1) DK3213752T3 (fr)
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WO (1) WO2016068600A1 (fr)

Families Citing this family (9)

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Publication number Priority date Publication date Assignee Title
KR101916652B1 (ko) * 2016-06-29 2018-11-08 올릭스 주식회사 작은 간섭 rna의 rna 간섭효과 증진용 화합물 및 이의 용도
KR20190128806A (ko) * 2018-05-09 2019-11-19 주식회사 하임바이오 암의 예방 또는 치료용 약학적 조성물
KR102141971B1 (ko) * 2018-11-12 2020-08-06 주식회사 노암 항암용 조성물
KR102127125B1 (ko) * 2018-12-31 2020-06-26 주식회사 노암 면역체크포인트 억제제를 포함하는 항암용 조성물
WO2021020902A1 (fr) * 2019-07-30 2021-02-04 주식회사 하임바이오 Nouveau composé anticancéreux et composition pharmaceutique le comprenant en tant que principe actif
KR20220008525A (ko) 2020-07-14 2022-01-21 주식회사 바이오파마 벤질티오 아르테미시닌을 유효성분으로 함유하는 대장암 세포 및 대장암 줄기세포 성장 억제용 약학 조성물
KR102562739B1 (ko) * 2020-09-11 2023-08-03 연세대학교 산학협력단 암의 기원 세포의 사멸용 약학적 조성물
KR20220036113A (ko) 2020-09-15 2022-03-22 주식회사 바이오파마 신규한 페녹시아르테미시닌을 유효성분으로 함유하는 대장암 세포 및 대장암 줄기세포 성장 억제용 약학 조성물
KR20220120836A (ko) 2021-02-24 2022-08-31 주식회사 바이오파마 알킬티오아르테미시닌을 함유하는 대장암 세포 및 대장암 줄기세포 성장 억제용 약학 조성물

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20130004770A (ko) * 2011-07-04 2013-01-14 한양대학교 산학협력단 암 줄기세포 성장 억제용 조성물
US20140065246A1 (en) * 2003-05-30 2014-03-06 Charles E. Zeilig Intracellular calcium modulation for cancer treatment
US8685926B2 (en) * 2007-09-26 2014-04-01 The Regents Of The University Of Colorado, A Body Corporate Compositions and method for CaMKII inhibitors and uses thereof

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002028886A2 (fr) * 2000-10-05 2002-04-11 Mcmaster University Caloxines
US20110098232A1 (en) * 2003-05-30 2011-04-28 Zeilig Charles E Methods For The Selective Treatment Of Tumors By Calcium-Mediated Induction Of Apoptosis
KR101900478B1 (ko) * 2011-08-04 2018-09-20 한올바이오파마주식회사 암의 재발 또는 전이의 억제용 약제학적 조성물
ES2902484T3 (es) * 2014-05-16 2022-03-28 Univ Yale Composiciones para el tratamiento y la prevención de la pancreatitis y las lesiones renales

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140065246A1 (en) * 2003-05-30 2014-03-06 Charles E. Zeilig Intracellular calcium modulation for cancer treatment
US8685926B2 (en) * 2007-09-26 2014-04-01 The Regents Of The University Of Colorado, A Body Corporate Compositions and method for CaMKII inhibitors and uses thereof
KR20130004770A (ko) * 2011-07-04 2013-01-14 한양대학교 산학협력단 암 줄기세포 성장 억제용 조성물

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CAO, X. ET AL.: "Glucose uptake inhibitor sensitizes cancer cells to daunorubicin and overcomes drug resistance in hypoxia", CANCER CHEMOTHERAPY AND PHARMACOLOGY, vol. 59, 2007, pages 495 - 505, XP019472993 *
MUELLER, M. ET AL.: "Combined targeted treatment to eliminate tumorigenic cancer stem cells in human pancreatic cancer", GASTROENTEROLOGY, vol. 137, no. 3, 2009, pages 1102 - 1113, XP026793002, DOI: doi:10.1053/j.gastro.2009.05.053 *

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US20170312333A1 (en) 2017-11-02
ES2743493T3 (es) 2020-02-19
CN107106580A (zh) 2017-08-29
CN107106580B (zh) 2020-12-04
KR20170083997A (ko) 2017-07-19
EP3213752A4 (fr) 2018-07-25
EP3213752A1 (fr) 2017-09-06
KR101774652B1 (ko) 2017-09-05
KR20160051638A (ko) 2016-05-11
DK3213752T3 (da) 2019-09-02
EP3213752B1 (fr) 2019-08-14
JP2017534628A (ja) 2017-11-24
JP6446134B2 (ja) 2018-12-26
US10420814B2 (en) 2019-09-24

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